title: 4C-seq_PromoterViewpoint_etoh_dex_rep1
description: Protocols: 4C-seq was performed with 5 million cells per treatment, cells were grown in DMEM containing 5% fetal bovine serum. A549 cells were grown to approximately 90% confluence according to standard protocols in DMEM medium with 5% FBS. Cells were treated with either 100 nM dexamethasone or 0.1% ethanol. After 4 hours, cells were washed 2x with PBS and cultured in hormone-free medium for 24 hours, after which cells were treated again for 4 hours with either 100 nM dexamethasone or 0.1% ethanol. 4C template preparation was carried out as described in PMID: 22929766. Restriction digests were performed with the enzymes Csp6I (primary, Thermo Fisher Scientific) and DpnII (secondary, Thermo Fisher Scientific). 4C-seq library preparation was performed as described in PMID: 31351925 (except that the DNA was purified with 1.5× AMPure XP beads (Beckman Coulter) after the first PCR). The following primer pairs were used for the inverse PCR: ZBTB16 promoter-proximal VP (fwd: TACACGACGCTCTTCCGATCTAGCAGAGAGGAGTTGAGG, rev: ACTGGAGTTCAGACGTGTGCTCTTCCGATCTTAGTGAACCAGGTGCCAG), ZBTB16 intronic VP (fwd: TACACGACGCTCTTCCGATCTATGTGTGCGTTCATGTATGT, rev: ACTGGAGTTCAGACGTGTGCTCTTCCGATCTGAGGAAAGGTTAGGAAGTGG)
attributes.tag: 1st restriction digest
attributes.value: Csp6I
attributes.tag: 2nd restriction digest
attributes.value: DpnII
attributes.tag: ENA first public
attributes.value: 2021-05-12
attributes.tag: ENA last update
attributes.value: 2021-05-12
attributes.tag: External Id
attributes.value: SAMEA7722616
attributes.tag: INSDC center name
attributes.value: MPIMG
attributes.tag: INSDC first public
attributes.value: 2021-05-03T00:22:16Z
attributes.tag: INSDC last update
attributes.value: 2020-12-15T12:54:48Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9915:4C-seq_PromoterViewpoint_etoh_dex_rep1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: A549
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: carcinoma
attributes.tag: organism part
attributes.value: lung
attributes.tag: sample name
attributes.value: E-MTAB-9915:4C-seq_PromoterViewpoint_etoh_dex_rep1
attributes.tag: viewpoint
attributes.value: chr11:113929670-113930464 (hg19)